Ruolo biologico dei fenilpropanoidi in colture cellulari di Daucus carota

Plants produce many thousands of secondary metabolites and to date from the chemical point of view more than 40,000 terpenoids, 5,000 alkaloids and 20,000 phenolic compounds are known. It is known that many biotic and abiotic stresses that induce the production of reactive oxygen species (ROS) in plant cells induce the accumulation of phenylpropanoids, but if they are really involved in the reactions of scavenging of ROS in vivo is still unclear (Hernandez et al., 2009). In vitro plant cell cultures are widely used as experimental systems for studying many physiological, biochemical and molecular features of plant. R3M carrot cell culture was selected from a non-pigmented culture for their ability to produce anthocyanins. Experiments of in vivo imaging of single cells showed that pigment cells are more resistant than non-pigmented ones to oxidative stress. The metabolic characterization of metanolic extracts of R3M cell cultures by HPLC-DA and HPLC-MS reveals that R3M cell line produces anthocyanins, hydroxycinnamic and hydroxybenzoic acids. The aim of this doctoral project is an investigation of the possible biological role of the presence and relative abundance of some phenylpropanoids, particularly anthocyanins, in R3M cell culture. The accumulation of anthocyanins has been increased by feeding of biosynthetic precursors. The HPLC-MS analysis of cell methanol extracts has shown the effect of feeding on the R3M metabolic profile; comparison of the treatments has been performed by multivariate statistical analysis of the chromatographic data. The different ability of feeded cells to respond to thermal stress has been analyzed by microscopic observation of phenotypic traits related to damage caused by heat and by assessing the degree of lipid peroxidation. The experiments have revealed important aspects of the role that some phenylpropanoids have in response to adverse environmental conditions in vivo.